•快速—纯化抗体所需的时间是蛋白 A 或 G 所需时间的一半 • 高效—回收率和纯度通常优于蛋白 A 或 G 纯化 • 稳健—对与蛋白 A 或 G 的结合能力较差的 IgG 亚类有效 • 温和—抗体可保留活性,因为它们不暴露于变性条件 • 可再生—支持物可再生,并可用于多达 6 次纯化 • 兼容—无胺缓冲液无需去除或中和操作 • 灵活—可容纳多种样品体积和处理容器;细胞培养物: 离心柱可用于纯化 < 30 mL 上清液;过滤瓶可用于批量处理多达 1 L 的样品。腹水:离心柱可用于纯化多达 6 mL
与典型的结合和洗脱亲和方法不同,Melon 凝胶纯化基于所制备的上清液或腹水样品中的非 IgG 蛋白结合,从而能够在流过物(非结合)组分中回收纯 IgG。回收的抗体可立即用于测定或后续偶联和其他修饰步骤。无需通过中和或脱盐来降低苛刻洗脱缓冲液的影响。与传统蛋白 A 或蛋白 G 方法相比,Melon 凝胶纯化可节约时间,且不影响抗体的质量。
足够用于:从多达 1 L 细胞培养物上清液或多达 200 mL 腹水中纯化 IgG • Melon 凝胶支持物,200mL • Melon 凝胶纯化缓冲液(干燥混合物,可制备 100X 制剂),1 L • Melon 凝胶再生剂(干燥混合物,可制备 5X 制剂),1 L
储存在 4°C 下
常见问题解答 (FAQ)
How can I determine when the Melon Gel Support is saturated?
For serum, a volume double the Melon Gel Support volume is the maximum amount that can be processed at one time. To determine if too much sample was processed, examine the purified IgG by SDS-PAGE for additional protein bands.
No. Thermo Scientific Thiophilic Adsorbent (Cat. Nos. 20500 and 44916) can be used to purify IgY from serum. The Chicken IgY Purification Kit (Cat. No. 44918) can be used to purify IgY from egg yolks.
Is the Melon Gel IgG Spin Purification Kit (Cat. No. 45206) a viable alternative to using traditional Protein A and Protein G methods for purification of IgG from serum?
Yes. With the Melon Gel IgG Spin Purification Kit, 90% recovery of IgG with greater than 80% purity are typically observed for a single purification. Although IgA and IgM may be present along with the purified IgG, a high percentage (> 80%) of IgA and IgM is retained on the gel support. This high binding of IgA and IgM to the gel support combined with the fact that IgA and IgM are present in low abundance in serum results in low levels of these antibody molecules in the purified IgG. Here are other advantages of using the Melon Gel IgG Spin Purification Kit over traditional Protein A or Protein G methods:
-Simple, one-step protocol - IgG purification with Melon Gel Resin does not involve binding, washing, and multiple elution steps required by Protein A and Protein G
-Rapid purification - antibody purification from serum is four to six times faster than Protein A or G methods
-Minimal dilution - reduced need to concentrate antibody before storage or use
-Robust purification - Melon Gel IgG Purification Kits work with a wide range of antibodies including many that do not purify well on Protein A or Protein G
-Gentle purification - No harsh antibody elution conditions helps retain IgG activity
-Regenerable support - Melon Gel Resin can be used for multiple antibody purifications
The Melon Gel Support can be used up to three times. Performing an ammonium sulfate precipitation on serum samples containing large amounts of non-antibody protein before purification improves purification efficiency and increases reusability.